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Epithelial to mesenchymal transition (EMT) in human prostate cancer: lessons learned from ARCaP model

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Abstract

Androgen refractory cancer of the prostate (ARCaP) cells contain androgen receptor (AR) and synthesize and secrete prostate specific antigen (PSA). We isolated epithelia-like ARCaPE from parental ARCaP cells and induced them to undergo epithelial–mesenchymal transition (EMT) by exposing these cells to soluble factors including TGFβ1 plus EGF, IGF-1, β2-microglobulin (β2-m), or a bone microenvironment. The molecular and behavioral characteristics of the resultant ARCaPM were characterized extensively in comparison to the parental ARCaPE cells. In addition to expressing mesenchymal biomarkers, ARCaPM gained 100% incidence of bone metastasis. ARCaPM cells express receptor activator of NF-κB ligand (RANKL), which was shown to increase tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in culture, and when metastatic to bone in vivo. We provide evidence that RANKL expression was promoted by increased cell signaling mediated by the activation of Stat3-Snail-LIV-1. RANKL expressed by ARCaPM cells is functional both in vitro and in vivo. The lesson we learned from the ARCaP model of EMT is that activation of a specific cell signaling pathway by soluble factors can lead to increased bone turnover, mediated by enhanced RANKL expression by tumor cells, which is implicated in the high incidence of prostate cancer bone colonization. The ARCaP EMT model is highly attractive for developing new therapeutic agents to treat prostate cancer bone metastasis.

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Abbreviations

AR:

Androgen receptor

ARCaP:

Androgen-refractory human prostate cancer cell model from a patient with prostate cancer bone metastasis

ARCaPE :

ARCaP clone with epithelial phenotype

ARCaPM :

ARCaP clone with mesenchymal phenotype

β2-m:

β2-Microglobulin

BSP:

Bone sialoprotein

C4-2:

Lineage derivative cells from LNCaP

C4-2B:

C4-2 cells metastasized to bone

CK18/19:

Cytokeratin 18/19

CM:

Conditioned medium

CREB:

cAMP-responsive element-binding protein

E-cad:

E-cadherin

EGF:

Epidermal growth factor

EMT:

Epithelial–mesenchymal transition

FBS:

Fetal bovine serum

IC:

Intracardiac

IGF-1:

Insulin-like growth factor 1

IHC:

Immunohistochemistry

IL13Rα2:

Interleukin13 receptor α2

LNCaP:

Prostate cancer cells metastasized to lymph node

MET:

Mesenchymal–epithelial transition

MErT:

Mesenchymal–epithelial-reverting transition

MMT:

Mesenchymal–mesenchymal transition

N-cad:

N-cadherin

NFκB:

Nuclear factor kappa B

OC:

Osteocalcin

OPG:

Osteoprotegerin

OPN:

Osteopontin

RANKL:

Receptor activator of NFκB ligand

RT-PCR:

Reverse transcriptase-polymerase chain reaction

SCID:

Severe combined immunodeficiency

siRNA:

Small interfering RNA

Stat:

Signal transducer and activator of transcription

TGFβ1:

Transforming growth factor β1

TRAP:

Tartrate-resistant acid phosphatase

VM:

Vimentin

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Acknowledgements

We thank Gary Mawyer for editing. The helpful discussion and support from Guodong Zhu, Weiping Qian, Daqing Wu and Clayton Yates are greatly appreciated. The authors are particularly indebted to the helpful suggestions made by the editors, Drs. Rik Thompson and Elizabeth Williams, during the revision of this manuscript. This study was supported in part by CA082739 (HYEZ), U54 CA119338, CA098912 and CA766201 (LWKC). We are grateful to the generous gift from Frances and Clarence Wilkins in support of this study.

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Authors and Affiliations

  1. Molecular Urology & Therapeutics Program, Department of Urology, Emory University School of Medicine, 1365B Clifton Road, Suite 5107, Atlanta, GA, 30322, USA

    Haiyen E. Zhau, Valerie Odero-Marah, Hui-Wen Lue, Takeo Nomura, Ruoxiang Wang, Gina Chu, Wen-Chin Huang & Leland W. K. Chung

  2. Department of Biology, Georgia State University, Atlanta, GA, 30302, USA

    Hui-Wen Lue, Gina Chu & Zhi-Ren Liu

  3. Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch, Galveston, TX, 77555, USA

    Binhua P. Zhou

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  1. Haiyen E. Zhau
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Correspondence to Haiyen E. Zhau or Leland W. K. Chung.

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Zhau, H.E., Odero-Marah, V., Lue, HW. et al. Epithelial to mesenchymal transition (EMT) in human prostate cancer: lessons learned from ARCaP model. Clin Exp Metastasis 25, 601–610 (2008). https://doi.org/10.1007/s10585-008-9183-1

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  • DOI: https://doi.org/10.1007/s10585-008-9183-1

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